Composition with effects of decline prevention, improvement or enhancement of normal responses of cognitive abilities of a healthy person

ABSTRACT

Arachidonic acid and/or compounds containing arachidonic acid as a constituent fatty acid are used to obtain compounds which prevent decline of, improve or enhance normal responses of cognitive abilities of a healthy person, and which are highly suitable for pharmaceuticals as well as food products, with minimal side effects.

FIELD OF THE INVENTION

The present invention provides foods and beverages with effects ofdecline prevention, improvement or enhancement of normal responses ofcognitive abilities of a healthy person, containing, as activeingredients, arachidonic acid and/or compounds with arachidonic acid asa constituent fatty acid, and a process for their production. Morespecifically, the invention provides foods and beverages with effects ofdecline prevention, improvement or enhancement of normal responses ofcognitive abilities of a healthy person, awareness level and/ordiscriminatory ability with respect to events selected from the groupconsisting of auditory stimuli, visual stimuli, gustatory stimuli,olfactory stimuli and somatosensory stimuli, which contain, as activeingredients, one or more species selected from the group consisting ofarachidonic acid, arachidonic acid alcohol esters and triglycerides,phospholipids or glycolipids containing arachidonic acid as part or allof the constituent fatty acid, as well as a process for theirproduction.

BACKGROUND ART

Cognition is the process of selecting information from the externalenvironment and clearly identifying it, and more specifically, oftransmitting external stimuli such as visual, auditory, olfactory,gustatory and somatosensory stimuli to the brain through sensory organsand processing and precisely identifying them by coordinated functioningof multiple regions of the brain. Cognitive abilities consist of the twofactors of information processing speed (speed of transforming externalstimuli) and the amount of information processing resources (the levelof resource allocation for information processing on external stimuli),and the effects of decline prevention, improvement or enhancement ofnormal responses of cognitive abilities of a healthy person according tothe present invention are therefore understood to refer to declineprevention, improvement or enhancement of normal functions such asattention, memory, perception, language, calculation and the like, whilebeing distinct from improvement of impaired function.

Various therapeutic agents have been researched and developed inrelation to impaired cognitive function. Unfortunately, however, noeffective compounds have yet been found that prevent decline of, improveor enhance normal responses of cognitive abilities of a healthy person.That is, although methods are known for determining cognitive functionimpairment and evaluating improving effects (see, for example,Nishimura, T., Takeda, M., “Diagnostic imaging of Alzheimer's dementia”,pp. 27-36, Medical View Publications (Feb. 10, 2001)) are known, suchmethods do not allow evaluation of decline prevention, improvement orenhancement of normal responses. A need has therefore existed for aneffective means of objectively evaluating normal responses of cognitiveabilities of a healthy person in order to objectively develop andresearch drug agents for decline prevention, improvement or enhancementof normal responses of cognitive abilities of a healthy person.

Attention has recently become focused on event related potentialss ofbrain as a physiological index to serve as an objective evaluation ofcognitive ability. It is possible to record the electrical activityexhibited by neurons through electrodes attached on the human scalp.Background brain waves are generally known as continuous electricalactivity which varies depending on the arousal level. A transient minuteaction potential is also present which varies in relation to a givenspecifiable event. This action potential is currently referred to as the“event related potentialss of brain”, and is interpreted as thepotential induced by internal or external stimuli. The event relatedpotentialss of brain, a minute potential variation of about 0.1 μV toseveral tens of μV, is composed of a series of positive waves andnegative waves. The positive wave appearing near 300 msec for thestimulus event among this series of potential variation is associatedwith cognitive function and is termed P300. The time to appearance ofP300 for a stimulus event is called the P300 latency and representsinformation processing speed (speed of transforming external stimuli),while the height of the peak from the baseline is called the P300amplitude and represents the amount of information processing resources(the level of resource allocation for information processing on externalstimuli) (see, for example, Mitamura, S., “Current Therapy”, Vol. 18,No.4, pp. 618-621, Life Medicom (Feb. 10, 2001)). It has thus becomepossible to objectively measure human cognitive response by measuringthe event related potentialss of brain.

External stimuli are transmitted through sensory organs to the brain,and the brain is the tissue like adipose mass, about ⅓ of the whitematter and ¼ of the gray matter consists of phospholipids. Because mostof the polyunsaturated fatty acids of the phospholipids constituting thecell membranes of the brain are arachidonic acid and docosahexaenoicacid, it has been suggested that these polyunsaturated fatty acids mayplay some role in enhancing learning and memory abilities and preventingor improving senile dementia. Arachidonic acid and docosahexaenoic acidcannot be synthesized de novo in animals and therefore must be ingestedfrom food either directly or indirectly (as linoleic acid precursor ofarachidonic acid or α-linoleic acid precursor of docosahexaenoic acid).Attention has thus been focused on enhancing effects on learning andmemory and preventive and improving effects on senile dementia byexternally supplying arachidonic acid and docosahexaenoic acid.Docosahexaenoic acid is found abundantly in fish oil sources and a greatdeal of research has been conducted on its effects of brain functionimprovement, while inventions have been disclosed in regard to its usein learning ability reinforcers, memory reinforcers, dementia preventingagents, dementia treatment agents, anti-dementia agents or functionalfoods with brain function-improving effects (see, for example, JapaneseUnexamined Patent Publication No. 7-82146, Japanese Unexamined PatentPublication No. 5-117147 and Japanese Unexamined Patent Publication No.2-49723). Furthermore, arachidonic acid and/or compounds containingarachidonic acid as a constituent fatty acid have recently been shown bythe present inventors, to improve age-related decline in learningability, based on results of administering arachidonic acid and/or acompound comprising arachidonic acid as a constituent fatty acid to agedanimals subjected to a Morris water maze test, as described in JapaneseUnexamined Patent Publication No. 2003-48831, which was published onFeb. 21, 2003, entitled “Composition Having Effects Of Preventing OrAmeliorating Conditions Or Diseases Caused By Brain Hypofunction”.However, this particular invention is directed toward decline in brainfunction and discloses nothing regarding effects on normal responses ofcognitive abilities of a healthy person.

Some attempts have been made to determine the improving effects ofseveral compounds on cognitive response based on event relatedpotentialss of brain. Miyanaga, K. examined the pharmacological actionof DHA on brain function, measuring event related potentialsss of brainbefore and after orally administering capsules containing 2400 mg DHA to26 healthy subjects, and discovered that the P300 latency wassignificantly shortened and the P300 amplitude was significantlyaugmented (see Miyanaga, K., “Shoku no Kagaku” [Food Science] pp. 84-96,Korin Books (1999)). However, due to a lack of comparative results withplacebo samples, no correlation could be established between blood DHAlevels and P300 results. In a prolonged administration test with dailyadministration of capsules containing 900 mg DHA to 97 healthy geriatricsubjects for 6 months, no change in P300 was observed, thus leavinguncertainty about the efficacy of DHA.

Therefore, while docosahexaenoic acid is considered to exhibit animproving effect on learning ability, the fact that no effective resultswere found in terms of the event related potentials of brain as an indexof cognitive ability means that it remains unclear whether or notarachidonic acid and/or compounds with arachidonic acid as a constituentfatty acid are effective for decline prevention, improvement andenhancement of normal responses of cognitive abilities of a healthyperson.

DISCLOSURE OF THE INVENTION

Problems to be Solved by the Invention

Consequently, it has been ardently desired to develop compounds whichprevent decline of, improve or enhance normal responses of informationprocessing speed (speed of transforming external stimuli) and the amountof information processing resources (the level of resource allocationfor information processing on external stimuli), as the two factors ofcognitive abilities, and which are highly suitable for pharmaceuticalsas well as food products, with minimal side effects.

Means for Solving the Problems

The present invention therefore provides foods and beverages witheffects of decline prevention, improvement or enhancement of normalresponses of cognitive abilities of a healthy person, containing, asactive ingredients, arachidonic acid and/or compounds with arachidonicacid as a constituent fatty acid, and a process for their production.More specifically, the invention provides foods and beverages witheffects of decline prevention, improvement or enhancement of normalresponses of cognitive abilities of a healthy person, awareness leveland/or discriminatory ability with respect to events (auditory stimuli,visual stimuli, gustatory stimuli, olfactory stimuli, somatosensorystimuli), which containes as active ingredients one or more speciesselected from the group consisting of arachidonic acid, arachidonic acidalcohol esters and triglycerides, phospholipids or glycolipidscomprising arachidonic acid as part or all of the constituent fattyacid, as well as a process for their production.

As a result of diligent research for the purpose of elucidating theeffects of decline prevention, improvement or enhancement of normalresponses of cognitive abilities of a healthy person by arachidonic acidand/or compounds comprising arachidonic acid as a constituent fattyacid, the present inventors surprisingly discovered an effect ofarachidonic acid and/or compounds comprising arachidonic acid as aconstituent fatty acid, based on evaluation using the event relatedpotentials of brain as an index.

The present inventors also succeeded in achieving microbial industrialproduction of triglycerides containing at least 10 wt % arachidonicacid, and elucidated the effects of those triglycerides in effects testsaccording to the invention.

The present inventors further succeeded in achieving enzymaticproduction of oils and fats containing triglycerides with medium-chainfatty acids bound at the 1,3-positions and arachidonic acid bound at the2-position, and elucidated the effects of those triglycerides in effectstests according to the invention.

The present invention therefore provides foods and beverages witheffects of decline prevention, improvement or enhancement of normalresponses of cognitive abilities of a healthy person, containing asactive ingredients arachidonic acid and/or compounds with arachidonicacid as a constituent fatty acid, and a process for their production.More specifically, the invention provides foods and beverages witheffects of decline prevention, improvement or enhancement of normalresponses of cognitive abilities of a healthy person, awareness leveland/or discriminatory ability with respect to events (auditory stimuli,visual stimuli, gustatory stimuli, olfactory stimuli, somatosensorystimuli), which containes as active ingredients one or more speciesselected from the group consisting of arachidonic acid, arachidonic acidalcohol esters and triglycerides, phospholipids or glycolipidscomprising arachidonic acid as part or all of the constituent fattyacid, as well as a process for their production.

According to the invention it is possible to provide foods and beverageshaving effects of decline prevention, improvement or enhancement ofnormal responses of cognitive abilities of a healthy person whichcontaines as active ingredients arachidonic acid and/or compounds witharachidonic acid as a constituent fatty acid, and a process for theirproduction, thus the invention is particularly useful in modern society.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a bar graph showing changes in arachidonic acid levels inserum phospholipids before and after ingestion of arachidonicacid-containing edible oil capsules and olive oil-containing capsules(placebo capsules).

FIG. 2 is a line graph showing changes in P300 latency before and afteringestion of arachidonic acid-containing edible oil capsules and oliveoil-containing capsules (placebo capsules).

FIG. 3 is a line graph showing changes in P300 amplitude before andafter ingestion of arachidonic acid-containing edible oil capsules andolive oil-containing capsules (placebo capsules).

FIG. 4 is a pair of dot graphs showing correlation between P300 (P300latency and amplitude) and arachidonic acid levels in serumphospholipids.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to foods and beverages with effects ofdecline prevention, improvement or enhancement of normal responses ofcognitive abilities of a healthy person, comprising as activeingredients arachidonic acid and/or compounds with arachidonic acid as aconstituent fatty acid, and to a process for their production.

The compositions of the invention have effects of decline prevention,improvement or enhancement of normal responses of cognitive abilities ofa healthy person, or stated differently, they have effects of declineprevention, improvement or enhancement of normal responses of awarenesslevel of a healthy person and have effects of decline prevention,improvement or enhancement of normal responses of discriminatory abilityof a healthy person with respect to events (auditory stimuli, visualstimuli, gustatory stimuli, olfactory stimuli, somatosensory stimuli),and are able to exhibit their effects in foods or beverages,pharmaceuticals, quasi drugs and the like. The effects of declineprevention, improvement or enhancement of normal responses of cognitiveabilities of a healthy person are also effective as foods or beverages,pharmaceuticals and quasi drugs designed for the purpose of declineprevention, improvement or enhancement of normal functions such asattention, memory, perception, language, calculation and the like, andas foods or beverages, healthy foods, functional foods, special healthcare foods, infant foods, geriatric foods, etc. designed for the purposeof maintaining or enhancing concentration, maintaining attention,clearing the head or stimulating the mind, rejuvenation and the like.

The active ingredients according to the invention may be, instead offree arachidonic acid, any compounds having arachidonic acid as aconstituent fatty acid. Compounds having arachidonic acid as aconstituent fatty acid include arachidonic acid salts such as calciumand sodium salts, or arachidonic acid alcohol esters such as arachidonicmethyl ester, arachidonic acid ethyl ester and the like. Triglycerides,diglycerides, monoglycerides, phospholipids or glycolipids containingarachidonic acid as part or all of the constituent fatty acid may alsobe used.

For application to a food product, the arachidonic acid is preferably inthe form of triglycerides or phospholipids, and especiallytriglycerides. While virtually no natural sources of arachidonicacid-containing triglycerides (or triglycerides including triglycerideswith arachidonic acid as part or all of the constituent fatty acid) areknown, the present inventors have succeeded in enabling industrial useof triglycerides containing arachidonic acid as a constituent fattyacid, and have been the first to elucidate the effect of the activeingredient of the invention in humans by prolonged administration tohumans and analysis of event related potentials of brain allowingobjective evaluation of cognitive abilities, and to demonstrate that ithas an effect of decline prevention, improvement or enhancement ofnormal responses of cognitive abilities of a healthy person.

According to the present invention, therefore, there may be usedtriglycerides which include triglycerides wherein part or all of theconstituent fatty acids are arachidonic acid as the active ingredient ofthe invention (arachidonic acid-containing triglycerides). Forapplication to food products, the arachidonic acid-containingtriglycerides are preferably in the form oils or fats (triglycerides)wherein the proportion of arachidonic acid is at least 20 wt % (W/W),preferably at least 30 wt % and more preferably at least 40 wt % of thetotal fatty acids of the triglycerides. According to the invention,therefore, there may be used any arachidonic acid-containing oils orfats (triglycerides) obtained by culturing microbes capable of producingthe same.

As examples of microbes capable of producing arachidonic acid-containingoils or fats (triglycerides) there may be mentioned microbes belongingto the genus Mortierella, Conidiobolus, Pythium, Phytophthora,Penicillium, Cladosporium, Mucor, Fusarium, Aspergillus, Rhodotorula,Entomophthora, Echinosporangium or Saprolegnia. As microbes belonging tothe genus Mortierella subgenus Mortierella there may be mentionedMortierella elongata, Mortierella exigua, Mortierella hygrophila,Mortierella alpina, and the like. Specifically, there may be mentionedstrains such as Mortierella elongata IFO8570, Mortierella exiguaIFO8571, Mortierella hygrophila IFO5941, Mortierella alpina IFO8568,ATCC16266, ATCC32221, ATCC42430, CBS219.35, CBS224.37, CBS250.53,CBS343.66, CBS527.72, CBS529.72, CBS608.70, CBS754.68, etc.

These strains are all available without restriction from the Institutefor Fermentation, Osaka (IFO), the American Type Culture Collection(ATCC) and the Centralbureau voor Schimmelcultures (CBS). In addition,there may be used strain Mortierella elongata SAM0219 (FERM P-8703)(FERM BP-1239) separated from soil by the research group of the presentinventors.

For culturing of the strains used for the invention, the spores, hyphaor cells of a seed culture solution obtained by preculturing areinoculated to liquid or solid medium for main culturing. In the case ofa liquid medium, the carbon source used may be any commonly used onesuch as glucose, fructose, xylose, saccharose, maltose, solubilizedstarch, molasses, glycerol or mannitol, although there is no limitationto these. As nitrogen sources there may be used organic nitrogensources, including natural nitrogen sources such as peptone, yeastextract, malt extract, meat extract, casamino acid, corn steep liquor,soybean protein, defatted soybean, cottonseed meal or the like, as wellas urea, and inorganic nitrogen sources such as sodium nitrate, ammoniumnitrate and ammonium sulfate. If necessary, trace nutrient sourcesincluding inorganic salts such as phosphoric acid salts, magnesiumsulfate, iron sulfate and copper sulfate or vitamins may also be used.These medium components are not particularly restricted so long as theyare at concentrations which do not inhibit growth of the microbes. Inpractice, the carbon source will generally be added in a total amount of0.1-40 wt % (W/V) and preferably 1-25 wt % (W/V). The initial nitrogensource addition may be at 0.1-10 wt % (W/V) and preferably 0.1-6 wt %(W/V), with further addition of the nitrogen source during culturing ifdesired.

Oils or fats (triglycerides) containing at least 45 wt % (W/W)arachidonic acid may be prepared as the active ingredient for theinvention by controlling the carbon source concentration in the medium.The culturing produces cells in the growth phase up to day 2-4 ofculturing, and cells in the oil/fat accumulating phase after day 2-4 ofculturing. The initial carbon source concentration is 1-8 wt % andpreferably 1-4 wt %, and the carbon source is successively added onlyduring the initial periods of the cell growth phase and oil/fataccumulating phase, up to a total successive carbon source addition of2-20 wt % and preferably 5-15 wt %. The successive addition of thecarbon source during the initial periods of the cell growth phase andoil/fat accumulating phase is based on the initial nitrogen sourceconcentration, for a carbon source concentration of zero in the mediumfrom the 7th day of culturing, preferably from the 6th day of culturingand more preferably from the 4th day of culturing, to obtain oils orfats (triglycerides) comprising at least 45 wt % arachidonic acid, as anactive ingredient according to the invention.

The culturing temperature for the arachidonic acid-producing microbewill differ depending on the microbe, but may be 5-40° C. and preferably20-30° C., or the cells may be cultured at 20-30° C. for growth and theculturing continued at 5-20° C. to produce unsaturated fatty acids. Suchtemperature management can also be used to increase the proportion ofpolyunsaturated fatty acids among the produced fatty acids. The pH ofthe medium may be 4-10 and preferably 5-9, and the culturing method maybe submerged culturing, shake culturing, or stationary culturing. Theculturing will usually be conducted for 2-30 days, preferably for 5-20days and more preferably for 5-15 days.

As a means for increasing the proportion of arachidonic acid in thearachidonic acid-containing oils or fats (triglycerides) other thancontrolling the carbon source concentration in the medium, thearachidonic acid-containing oils or fats may be selectively hydrolyzedto obtain arachidonic acid-rich oils or fats. Lipases used for selectivehydrolysis are not specific to triglyceride positions and, as thehydrolytic activity is lower in proportion to the number of doublebonds, fatty acid ester bonds other than those of polyunsaturated fattyacids are hydrolyzed. Also, transesterification occurs between theproduced PUFA triglycerides, resulting in triglycerides with increasedpolyunsaturated fatty acids (see, for example, Enhancement ofArachidonic Acid: Selective Hydrolysis of a Single-Cell Oil fromMortierella with Candida cylindracea lipase”, J. Am. Oil Chem. Soc., 72,pp. 1323-1327, AOCS Press (1998)). Thus, arachidonic acid-rich oils orfats (triglycerides) obtained by selective hydrolysis of arachidonicacid-containing oils or fats may be used as active ingredients accordingto the invention. A higher proportion of arachidonic acid is preferredwith respect to the total fatty acids in arachidonic acid-containingoils or fats (triglycerides) of the invention in order to minimize theeffects of the other fatty acids, but there is no restriction to a highproportion and, in practice, the absolute amount of arachidonic acid canpose a problem for application to food products, although oils or fats(triglycerides) containing arachidonic acid at 10 wt % or more may beused in practice.

As triglycerides comprising arachidonic acid as part or all of theconstituent fatty acid according to the invention there may be usedtriglycerides with medium-chain fatty acids bound at the 1,3-positionsand arachidonic acid bound at the 2-position. There may also be usedoils or fats (triglycerides) comprising at least 5 mole percent,preferably at least 10 mole percent, more preferably at least 20 molepercent and most preferably at least 30 mole percent of triglycerideswith medium-chain fatty acids bound at the 1,3-positions and arachidonicacid bound at the 2-position. The medium-chain fatty acids bound at the1,3-positions of the triglycerides may be selected from among fattyacids of 6 to 12 carbons. As examples of fatty acids of 6 to 12 carbonsthere may be mentioned caprylic acid and capric acid, with1,3-capryloyl-2-arachidonoyl-glycerol (hereinafter referred to as “8A8”)being preferred.

Triglycerides with medium-chain fatty acids bound at the 1,3-positionsand arachidonic acid bound at the 2-position are most suitable as oilsor fats (triglycerides) for infants or elderly persons. Ingested oils orfats (triglycerides) are usually hydrolyzed by pancreatic lipases uponentering the small intestine, and since pancreatic lipases are1,3-position specific type, the 1,3-positions of the triglycerides arecleaved to produce two free fatty acid molecules while simultaneouslyproducing one 2-monoacylglyceride (MG) molecule. Because 2-MG has veryhigh bile acid solubility and is highly absorbable, 2-position fattyacids are generally considered to be better absorbed. Also, 2-MGexhibits surfactant action when dissolved in bile acids, and thusfunctions to increase the absorption of the free fatty acids. The freefatty acids and 2-MG then form bile acid complex micelles withcholesterol or phospholipids and are taken up by intestinal epithelialcells, resulting in resynthesis of triacylglycerols and finally releaseinto the lymph as chylomicrons. However, the fatty acid properties ofthe pancreatic lipases are highest for saturated fatty acids, whilearachidonic acid is cleaved less efficiently. An additional problem isthat pancreatic lipase activity is lower in infants and the elderly,such that the ideal oils or fats (triglycerides) are triglycerides withmedium-chain fatty acids bound at the 1,3-positions and arachidonic acidbound at the 2-position.

One specific production process for triglycerides with medium-chainfatty acids bound at the 1,3-positions and arachidonic acid bound at the2-position is a production process whereby lipase is allowed to act onlyon 1,3-ester bonds of triglycerides in the presence ofarachidonic-containing oils or fats (triglycerides) and medium-chainfatty acids.

The oil or fat (triglycerides) used as the starting material consists oftriglycerides with arachidonic acid as a constituent fatty acid, andwhen the proportion of arachidonic acid is high with respect to thetotal fatty acids of the triglycerides, a temperature of 30-50° C. andpreferably 40-50° C. is employed which is higher than the ordinaryenzyme reaction temperature of 20-30° C., in order to prevent reductionin reaction yield due to increase in unreacted oil or fat (thetriglyceride starting material and the triglycerides wherein only one ofthe 1,3-position fatty acids is a medium-chain fatty acid).

As lipases which act specifically on the 1,3-position ester bonds oftriglycerides there may be mentioned those produced by microbesbelonging to the genus Rhizopus, Rhizomucor, Aspergillus or the like, aswell as pig pancreatic lipases. Such lipases may be commerciallyavailable ones. Examples thereof include Rhizopus delemar lipase(Talipase, product of Tanabe Seiyaku), Rhizomucor miehei lipase(Ribozyme IM by Novo Nordisk Co., Ltd.) and Aspergillus niger lipase(Lipase A, product of Amano Enzyme Co., Ltd.), with no particularlimitation to these enzymes, as any 1,3-position specific type lipasesmay be used.

The form in which the lipase is used is preferably as an immobilizedlipase on an immobilizing support, in order to confer heat stability tothe enzyme as the reaction temperature is 30° C. or higher andpreferably 40° C. or higher for increased reaction efficiency. Highlyporous resins are used as immobilizing supports, and there may bementioned ion-exchange resin supports with pores of at least about 100angstroms, such as Dowex MARATHON WBA (trademark of Dow Chemical).

A 0.5 to 20 times weight of aqueous solution of the 1,3-positionspecific type lipase with respect to the immobilizing support issuspended therein, and a 2 to 5 times amount of cold acetone (forexample, −80° C.) with respect to the suspension is slowly added whilestirring to form a precipitate. The precipitate is dried under reducedpressure to prepare the immobilized enzyme. As a simpler method, a 0.05to 0.4 times amount of the 1,3-position specific type lipase withrespect to the immobilizing support is dissolved in a minimal amount ofwater, and then the solution is mixed with the immobilizing supportwhile stirring and the mixture is dried under reduced pressure toprepare the immobilized enzyme. This procedure results in immobilizationof about 90% of the lipase on the support but with absolutely noexhibited transesterification activity, and therefore the immobilizedenzyme may be activated and provided for production most efficiently bypretreatment in the substrate (raw material oil or fat and medium-chainfatty acids) containing 1 to 10 wt % (W/V) water, and preferably in thesubstrate containing 1 to 3 wt % water.

The amount of water added to the reaction system is extremely importantdepending on the type of enzyme, since transesterification will notproceed in the absence of water, while an excess of water causeshydrolysis and a reduced glyceride yield (with diglycerides andmonoglycerides being produced upon hydrolysis). In such cases, however,the immobilized enzyme activated by pretreatment may be used to lessenthe importance of the amount of water added to the reaction system,allowing transesterification reaction to occur efficiently even in asystem containing no water. Moreover, the type of enzyme agent may alsobe selected to allow omission of pretreatment.

Thus, by using a heat-resistant immobilized enzyme and increasing theenzyme reaction temperature, it is possible to efficiently producetriglycerides with medium-chain fatty acids bound at the 1,3-positionsand arachidonic acid bound at the 2-position without reduction inreaction efficiency, even in the case of arachidonic acid-containingoils or fats (triglycerides) with low reactivity for 1,3-positionspecific type lipases.

The process for production of foods and beverages with effects ofdecline prevention, improvement or enhancement of normal responses ofcognitive abilities of a healthy person employs arachidonic acid and/ora compound with arachidonic acid as a constituent fatty acid, eitheralone or in admixture with a food or beverage material containingessentially no, or only a trace amount of, arachidonic acid. Here,“trace amount” means that arachidonic acid is present in the food orbeverage material but in an amount which does not reach the daily intakeof arachidonic acid according to the invention, as described hereunder,if the food composition is ingested by an individual.

Oils and fats (triglycerides) containing arachidonic acid as part or allof the constituent fatty acid have unlimited possibilities for use asraw materials and additives in foods, beverages, pharmaceuticals andquasi drugs. Furthermore, there are no limitations on the purpose oramounts of their use.

As examples of food compositions there may be mentioned common foodproducts as well as functional foods, nutritional supplements, modifiedmilk for immature infants, modified milk for infants, infant foodproducts, maternal foods or geriatric foods. As examples of foodproducts containing oils or fats there may be mentioned natural foodswhich originally contain oils and fats, such as meat, fish and nuts,food products with oils and fats added during preparation, such assoups, food products using oils or fats as heating media, such asdonuts, oil or fat food products such as butter, processed food productswith oils or fats added during processing, such as cookies, or foodproducts sprayed or coated with oils or fats during final processing,such as hard biscuits. They may also be added to agricultural foods,fermented foods, livestock foods, marine foods or beverages whichcontain no oils or fats. They may also be in the form of functional foodproducts, pharmaceuticals or quasi drugs, and for example, in the formof enteral nutrients, powders, granules, lozenges, oral solutions,suspensions, emulsions, syrups or the like.

The composition of the invention may also contain, in addition to theactive ingredient of the invention, various carriers and additivescommonly used in foods and beverages, pharmaceuticals or quasi drugs. Inparticular it preferably contains antioxidants in order to preventoxidation of the active ingredient of the invention. As antioxidantsthere may be mentioned natural antioxidants such as tocopherols, flavonederivatives, phyllodulcin, kojic acid, gallic acid derivatives,catechins, fukiic acid, gossypol, pyrazine derivatives, sesamol,guaiaol, guaiac acid, p-coumaric acid, nordihydroguaiatic acid, sterols,terpenes, nucleic acid bases, carotenoids, lignans and the like, andsynthetic antioxidants such as ascorbic palmitic ester, ascorbic stearicester, butylhydroxyanisole (BHA), butylhydroxytoluene (BHT),mono-t-butylhydroquinone (TBHQ) and 4-hydroxymethyl-2,6-di-t-butylphenol(HMBP). As tocopherols there may be mentioned α-tocopherol,β-tocopherol, γ-tocopherol, δ-tocopherol, ε-tocopherol, ξ-tocopherol,η-tocopherol and tocopherol esters (acetic acid tocopherol, etc.). Asexamples of carotenoids there may be mentioned β-carotene, cantaxanthin,astaxanthin and the like.

As supports to be used in the composition of the invention in additionto the active ingredient of the invention, there may be mentionedvarious types of carrier supports, extenders, diluents, fillers,dispersing agents, excipients, binder solvents (for example, water,ethanol or vegetable oils), solubilizing aids, buffers, dissolutionaccelerators, gelling agents, suspending agents, wheat flour, riceflour, starch, corn starch, polysaccharides, milk protein, collagen,rice oil, lecithin and the like. As examples of additives there may bementioned vitamins, sweeteners, organic acids, coloring agents,flavorings, dehumidifying agents, fibers, electrolytes, minerals,nutrients, antioxidants, preservatives, aromatic agents, humidifiers,natural edible extracts, vegetable extracts and the like, although thereis no limitation to these.

The principal medicinal component of the arachidonic acid or thecompound with arachidonic acid as a constituent fatty acid isarachidonic acid. The daily intake of arachidonic acid from food isreported to be 0.14 g in the Kanto region, Japan and 0.19-0.20 g in theKansai region, Japan (see “Shishitsu Eiyogaku” [Lipid Nutrition] 4, ed.by the Japan Society for Lipid Nutrition Editing Committee, pp.73-82(1995) ISN 1343-4594 CODEN:SHEIFG), and therefore arachidonic acidmust be ingested in a corresponding amount or greater. Thus, the dailyintake of arachidonic acid or a compound with arachidonic acid as aconstituent fatty acid according to the invention for an adult (60 kgbody weight, for example) is 0.001-20 g, preferably 0.01-10 g, morepreferably 0.05-5 g and most preferably 0.1-2 g, in terms of arachidonicacid.

When the active ingredient of the invention is actually used in a foodor beverage product, the absolute amount of arachidonic acid in the foodproduct is also important. However, since the absolute amount in thefood or beverage also varies depending on the amount of ingestion of thefood or beverage, the food product preferably contains the triglyceridesincluding triglycerides with arachidonic acid as part or all of theconstituent fatty acid, in an amount of at least 0.0003 wt %, preferablyat least 0.003 wt % and more preferably at least 0.03 wt % in terms ofarachidonic acid. When triglycerides with medium-chain fatty acids boundat the 1,3-positions and arachidonic acid bound at the 2-position areadded to a food or beverage, the amount of the triglycerides withmedium-chain fatty acids bound at the 1,3-positions and arachidonic acidbound at the 2-position is at least 0.001 wt %, preferably at least 0.01wt % and more preferably at least 0.1 wt %.

When the composition of the invention is used as a pharmaceuticalproduct, it may be produced by a common method for drug manufacturing,for example, a method described the Japanese Pharmacopoeia, or a methodbased thereon.

When the composition of the invention is used as a pharmaceuticalproduct, the content of the active ingredient in the composition is notparticularly restricted so long as the object of the invention isachieved, and the mixing proportion may be as deemed appropriate.

When the composition of the invention is used as a pharmaceuticalproduct, it is preferably administered in unit dosages, with oraladministration being particularly preferred. The doses of thecomposition of the invention will differ depending on the age, bodyweight and symptoms of the patient and the number of times administered,and for example, the arachidonic acid and/or compound with arachidonicacid as a constituent fatty acid according to the invention may usuallybe administered at about 0.001-20 g, preferably about 0.01-10 g, morepreferably about 0.05-5 g and most preferably about 0.1-2 g in terms ofarachidonic acid per day for an adult (approximately 60 kg), in divideddoses of 1 to 3 times a day.

The major fatty acids of brain phospholipid membranes are arachidonicacid and docosahexaenoic acid, and therefore the composition of theinvention is preferably a combination of arachidonic acid anddocosahexaenoic acid in consideration of maintaining balance. Generallyspeaking, arachidonic acid (n-6 unsaturated fatty acid) anddocosahexaenoic acid (n-3 unsaturated fatty acid) are biosynthesized bythe same enzyme from linoleic acid and α-linolenic acid, respectively.When arachidonic acid is administered alone, therefore, biosynthesis ofdocosahexaenoic acid is inhibited. Conversely, when docosahexaenoic acidis administered alone, biosynthesis of arachidonic acid is inhibited. Inorder to avoid such unbalances, it is preferred for arachidonic acid anddocosahexaenoic acid to be consumed in combination. Furthermore, as theproportion of eicosapentaenoic acid is very low in brain phospholipidmembranes, the combination preferably contains virtually noeicosapentaenoic acid. It therefore preferably contains noeicosapentaenoic acid, or only up to 1%. The composition more preferablycontains virtually no eicosapentaenoic acid with the arachidonic acidand docosahexaenoic acid. The combination of arachidonic acid anddocosahexaenoic acid also preferably has an arachidonicacid/docosahexaenoic acid ratio (by weight) in the range of 0.1-15 andpreferably in the range of 0.25-10. Most preferred are foods andbeverages containing eicosapentaenoic acid at no more than ⅕ (by weight)of the arachidonic acid.

The food compositions of the invention for foods or beverages, healthyfoods, functional foods, special health care foods, infant foods,geriatric foods and the like include those which are marketed usingpackaging containers and/or food composition merchandising tools (forexample, pamphlets or the like) which state or otherwise indicate thatthe compositions and/or components in the composition have effects ofdecline prevention, improvement or enhancement of cognitive abilities,or stated differently, that they have effects of decline prevention,improvement or enhancement of normal responses of awareness level of ahealthy person, effects of decline prevention, improvement orenhancement of normal responses of discriminatory ability of a healthyperson with respect to events (auditory stimuli, visual stimuli,gustatory stimuli, olfactory stimuli, somatosensory stimuli) and effectsof decline prevention, improvement or enhancement of normal functionssuch as attention, memory, perception, language, calculation and thelike, as well as having effects of maintaining or enhancingconcentration, maintaining attention, clearing the head or stimulatingthe mind, rejuvenation and the like.

EXAMPLES

The present invention will now be explained in greater detail throughthe following examples, with the understanding that these examples in noway limit the invention.

Example 1

Production Process for Triglycerides with Arachidonic Acid as aConstituent Fatty Acid

Mortierella alpina was used as an arachidonic acid-producing strain. A 6kL portion of medium containing 1.8% glucose, 3.1% defatted soybeanflour, 0.1% soybean oil, 0.3% KH₂PO₄, 0.1% Na₂SO₄, 0.05% CaCl₂.2H₂O and0.05% MgCl₂.6H₂O was prepared in a 10 kL culturing vessel, and theinitial pH was adjusted to 6.0. After seeding 30 L of the culturesolution, submerged culturing was conducted for 8 days under conditionsof 26° C. temperature, 360 m³/h aeration and 200 kPa interior pressure.The spinning was adjusted to maintain a dissolved oxygen concentrationof 10-15 ppm. Also, the glucose concentration of the medium was adjustedby feeding to be in the range of 1-2.5% up to the 4th day, and 0.5-1%thereafter (where the percentages are based on weight (W/V)). Aftercompletion of the culturing, followed by filtering and drying, the cellscontaining triglycerides with arachidonic acid as a constituent fattyacid were collected, the oil/fat was obtained from the obtained cells byhexane extraction, and the edible oil/fat was subjected to apurification step (degumming, deacidification, steam distilation,decoloration) to obtain 150 kg of arachidonic acid-containingtriglycerides (with arachidonic acid bound at any positions of thetriglycerides). The obtained oil/fat (triglyceride) was methylesterifiedand the resulting fatty acid methyl ester was analyzed by gaschromatography, which revealed an arachidonic acid proportion of 40.84%of the total fatty acids. The palmitic acid, stearic acid, oleic acid,linoleic acid, γ-linolenic acid and dihomo-γγ-linolenic acid contentswere 11.63%, 7.45%, 7.73%, 9.14%, 2.23% and 3.27%, respectively. Thearachidonic acid-containing oil/fat (triglyceride) was ethylesterifiedand 99% arachidonic acid ethyl ester was separated and purified from thefatty acid ethyl ester mixture comprising 40% arachidonic acid ethylester, by ordinary high performance liquid chromatography.

Example 2

Production of Triglycerides Including at Least 5% of Triglycerides withMedium-Chain Fatty Acids Bound at the 1,3-Positions and Arachidonic AcidBound at the 2-Position (8A8)

After suspending 100 g of an ion-exchange resin support (Dowex MARATHONWBA™: Dow Chemical) in 80 ml of 12.5% aqueous Rhizopus delemar lipasesolution (Talipase powder, product of Tanabe Seiyaku), the suspensionwas dried under reduced pressure to obtain the immobilized lipase.

Next, 80 g of the triglyceride containing 40 wt % arachidonic acidobtained in Example 1 (TGA40S), 160 g of caprylic acid, 12 g of theaforementioned immobilized lipase and 4.8 ml of water were reacted at30° C. for 48 hours while stirring (130 rpm). After completion of thereaction, the reaction mixture was removed to obtain the activatedimmobilized lipase.

A 10 g portion of the immobilized enzyme (Rhizopus delemar lipase,support: Dowex MARATHON WBA™) was packed into a jacketed glass column(1.8×12.5 cm, 31.8 ml volume), and a mixed oil containing the TGA40Sobtained in Example 1 and caprylic acid in a proportion of 1:2 waspassed through the column at a fixed flow rate (4 ml/h) for continuousreaction to obtain 400 g of reacted oil/fat. The column temperature was40-41° C. The unreacted caprylic acid and free fatty acids were removedfrom the reacted oil/fat by molecular distillation, and the edibleoil/fat was subjected to a purification step (degumming,deacidification, steam distillation, decoloration) to obtain8A8-containing oil/fat (triglycerides). The 8A8 content of the obtained8A8-containing oil/fat (triglycerides) was 31.6% as determined by gaschromatography and high performance liquid chromatography. (Theproportions of 8P8, 808, 8L8, 8G8 and 8D8 were 0.6, 7.9, 15.1, 5.2 and4.8%, respectively. The fatty acids P, O, L, G and D bound at thetriglyceride 2-position represent palmitic acid, oleic acid, linoleicacid, γ-linolenic acid and dihomo-γ-linolenic acid, respectively, 8P8 is1,3-capryloyl-2-palmitoleyl-glycerol, 808 is1,3-capryloyl-2-oleoyl-glycerol, 8L8 is1,3-capryloyl-2-linoleoyl-glycerol, 8G8 is1,3-capryloyl-2-γ-linolenoyl-glycerol and 8D8 is1,3-capryloyl-2-dihomo-γ-linolenoyl-glycerol.) Separation andpurification by high performance liquid chromatography according to anestablished protocol yielded 96 mole percent 8A8 from the obtained8A8-containing oil/fat (triglycerides).

Example 3

Production of Test Capsules

Water was added to 100 parts by weight of gelatin and 35 parts by weightof food grade glycerin for dissolution at 50-60° C. to prepare a gelatincoating with a viscosity of 2000 cp. Next, 0.05 wt % vitamin E oil wasmixed with the arachidonic acid-containing oil/fat (triglycerides)obtained in Example 1 to prepare Filling Content 1. Separately, 0.05 wt% vitamin E oil was mixed with oil/fat (triglycerides) containing 32mole percent of the 8A8 obtained in Example 2 to prepare Filling Content2. Filling Content 1 was used for molding and drying of capsules by acommon method to manufacture soft capsules containing 200 mg of fillingper capsule (arachidonic acid-containing edible oil capsules) whileFilling Content 2 was also used for molding and drying of capsules by acommon method to manufacture soft capsules containing 200 mg of fillingper capsule (8A8-containing edible oil capsules). Soft capsulescontaining olive oil as the filling content were also manufactured asplacebo capsules for a human test.

Example 4

Arachidonic Acid-Containing Edible Oil Capsule Ingestion Test toDetermine Effects on Cognitive Responses in Healthy Individuals

The event related potentials of brain (ERP) was measured by audible (orauditory discrimination) tasks according to the evoked potentialmeasuring guidelines established by the Evoked Potential ExaminationStandards Committee of the Japan Society of Clinical Neurophysiology.Specifically, an auditory oddball paradigm was utilized for the ERPmeasurement and both ears of the test subject were stimulated by twodifferent pure tones at frequencies of 1000 Hz and 2000 Hz were throughheadphones at ratio of 1:4 and in random order, and the subject wasinstructed to press a provided button when the 2000 Hz tone was heardand to count the number of times heard. The intensity of the tone was 90dB, the duration was 100 msec and the inter stimulus interval was randombetween 1000-3000 msec. The approximate number of stimulations in theactual test was 200 per trial and the duration of the test was about 10minutes. The test was conducted in 2 trials and the event relatedpotentialss of brain were measured. Electroencephalographies (EEGs) wererecorded by using Ag—AgCl electrodes, of which resistance is no greaterthan 5 kΩ, placed at three points along the mid-line of the scalp (Fz,Cz and Pz, according as international 10-20 displacements), andreference electrodes on both earlobes.

The maximum positive potential between 250-600 msec from the start ofthe low-frequency auditory simulation to be perceived (a 2000 Hz tone inthis experiment) was identified as the intrinsic component P300 whichvaries in evaluation of selective attention or cognitive response, thetime from the start of stimulation was recorded as the P300 latency(stimulation transmission speed) and the height of the potential fromthe baseline was recorded as the P300 amplitude (the amount ofinformation processing resources).

The human test for the invention was conducted with due consideration toadhering to the spirit of the Helsinki Declaration.

Upon explanation for consent to test participation, 12 consentinghealthy individuals (taking no medication, with no abnormal blood testresults and with no infarctions as determined by cranial CT scan) weredivided into two groups A and B (A: n=7, B: n=5). During a one monthperiod, Group A was administered three of the arachidonicacid-containing edible oil capsules prepared in Example 3 (80 mg/capsulearachidonic acid) for a daily arachidonic acid intake of 240 mg, whileGroup B was administered three placebo capsules. The event relatedpotentialss of brain were measured before and after capsule ingestion,and the P300 latency and amplitude were analyzed. The participants inGroups A and B were then taken off the capsules for a washout period ofone month. After the washout period, Group A was administered theplacebo capsules and Group B was administered the arachidonicacid-containing edible oil capsules for one month, and the event relatedpotentialss of brain were measured in the same manner before and aftercapsule ingestion (double blind, crossover test).

Blood was taken at the time of event related potentials of brainmeasurement, and the total lipids were extracted from the serum of eachparticipant by the Folch method. The lipids were fractionated bythin-layer chromatography, the phospholipid fraction was collected, thewater was removed by azeotropic distillation with ethanol, and analysiswas performed by gas chromatography upon conversion to fatty acid methylester with 10% HCl-methanol, to determine the arachidonic acid contentof the serum phospholipids.

FIG. 1 shows changes in arachidonic acid contents in serum phospholipidsbefore and after capsule ingestion. The arachidonic acid contents of theserum phospholipids of subjects ingesting the arachidonicacid-containing edible oil capsules were significantly increased afteringestion of the arachidonic acid-containing edible oil capsules,whereas the arachidonic acid contents of the serum phospholipids ofsubjects ingesting the placebo capsules were unchanged before and afteringestion of the placebo capsules.

FIG. 2 and FIG. 3 show changes in P300 latency and amplitude before andafter capsule ingestion. The P300 latency was significantly shortened by12.3 msec and the P300 amplitude was significantly increased by 1.9 μvwith ingestion of the arachidonic acid-containing edible oil capsules ascompared to the placebo capsules. A normal shortening in P300 latency of1.8 msec/year and decrease in P300 amplitude of 0.2 μV/year are known(Goodin D S et al. 1978), and therefore the results of this testindicate an average rejuvenation for cognitive responses of the subjectsof 6.8 years based on P300 latency and 9.5 years based on P300amplitude.

Next, the correlation between P300 (P300 latency and P300 amplitude) andserum arachidonic acid level (phospholipid) was determined by afirst-order curve based on the least square method using total of 48data obtained by measuring each of 12 subjects 4 times (FIG. 4). ForP300 latency, a significant correlation (coefficient of correlationR=−0.27) was found between the arachidonic acid levels, with shorterlatencies resulting from increased arachidonic acid levels. For P300amplitude as well, a significant correlation (coefficient of correlationR=−0.49) was found between the arachidonic acid levels, with larger P300amplitudes resulting from increased arachidonic acid levels. Thisconstitutes the first demonstration of improvement of cognitive responseby ingestion of arachidonic acid-containing edible oil, and the firstproof of arachidonic acid as the cause of the effect.

Example 5

8A8-Containing Edible Oil Capsule Ingestion Test to Determine Effects onCognitive Responses in Healthy Individuals

Upon explanation for consent to test participation in the same manner asExample 4, 16 consenting healthy individuals (taking no medication, withno abnormal blood test results and with no infarctions as determined bycranial CT scan) were divided into two groups A and B (n=8 for eachgroup). During a one month period, Group A was administered three of the8A8-containing edible oil capsules prepared in Example 3 (72 mg/capsulearachidonic acid) while Group B was administered three placebo capsules,and upon measuring the event related potentialss of brain and recordingthe latency and amplitude before and after capsule ingestion(double-blind test), the P300 latency was found to be significantlyshortened by 16.3 msec and the P300 amplitude significantly increased by2.4 μV due to ingestion of the 8A8-containing edible oil capsules. Theseresults indicate an average rejuvenation for cognitive responses of thesubjects of 9.1 years based on P300 latency and 12.0 years based on P300amplitude.

Example 6

Preparation of Capsules Containing Oil/Fat (Triglycerides) withArachidonic Acid as a Constituent Fatty Acid

Water was added to 100 parts by weight of gelatin and 35 parts by weightof food grade glycerin for dissolution at 50-60° C. to prepare a gelatincoating with a viscosity of 2000 cp. Next, 50 wt % of the arachidonicacid-containing oil/fat (triglycerides) obtained in Example 1 was mixedwith 50 wt % fish oil (tuna oil comprising eicosapentaenoic acid anddocosahexaenoic acid in proportions of 5.1% and 26.5%, respectively, ofthe total fatty acids), and this was combined with 0.05 wt % vitamin Eoil to prepare Filling Content 3. Separately, 80 wt % of the arachidonicacid-containing oil/fat (triglycerides) was mixed with 20 wt % fish oil(tuna oil comprising eicosapentaenoic acid and docosahexaenoic acid inproportions of 5.1% and 26.5%, respectively, of the total fatty acids),and this was combined with 0.05 wt % vitamin E oil to prepare FillingContent 4. The 99% arachidonic acid ethyl ester obtained in Example 1was also combined with 0.05 wt % vitamin E oil to obtain Filling Content5. Filling Contents 3-5 were used for molding and drying of capsules bya common method to manufacture soft capsules containing 200 mg offilling per capsule.

Example 7

Use in Oil Infusion Solutions

After combining 400 g of the oil/fat (triglycerides) containing 96% 8A8obtained in Example 2, 48 g of purified egg yolk lecithin, 20 g of oleicacid, 100 g of glycerin and 40 ml of 0.1 N caustic soda and dispersingthe mixture with a homogenizer, distilled water for injection was addedto 4 liters. This was emulsified with a high-pressure spray-typeemulsifier to prepare a lipid emulsion. The lipid emulsion was dispensedinto plastic bags in portions of 200 ml and subjected to high-pressuresteam sterilization treatment at 121° C. for 20 minutes to prepare oilinfusion solutions.

Example 8 Use in Juice

A 2 g portion of β-cyclodextrin was added to 20 ml of a 20% aqueousethanol solution, and then 100 mg of the arachidonic acid-containingtriglycerides obtained in Example 1 (comprising 0.05% vitamin E) wasadded thereto while stirring with a stirrer, and the mixture wasincubated at 50° C. for 2 hours. After cooling to room temperature(approximately 1 hour), incubation was continued at 4° C. for 10 hoursunder continuous stirring. The produced precipitate was recovered bycentrifugal separation, and after washing with n-hexane, it waslyophilized to obtain 1.8 g of a cyclodextrin clathrated compoundcontaining arachidonic acid-containing triglycerides. A 1 g portion ofthis powder was uniformly mixed with 10 L of juice to prepare juicecontaining arachidonic acid-containing triglycerides.

1. A composition with effects of decline prevention, improvement orenhancement of normal responses of cognitive abilities of a healthyperson, containing arachidonic acid and/or a compound with arachidonicacid as a constituent fatty acid.
 2. A composition according to claim 1,wherein the compound with arachidonic acid as a constituent fatty acidis an arachidonic acid alcohol ester or a triglyceride, phospholipid orglycolipid containing arachidonic acid as part or all of the constituentfatty acid.
 3. A composition according to claim 2, wherein thetriglyceride containing arachidonic acid as part or all of theconstituent fatty acid is a triglyceride having medium-chain fatty acidsbound at the 1,3-positions and arachidonic acid bound at the 2-position.4. A composition according to claim 3, wherein the medium-chain fattyacids are selected from among fatty acids of 6 to 12 carbons.
 5. Acomposition according to claim 4, wherein the medium-chain fatty acidsare selected from among fatty acids of 8 carbons.
 6. A composition witheffects of decline prevention, improvement or enhancement of normalresponses of cognitive abilities of a healthy person, which containstriglycerides including triglycerides containing arachidonic acid aspart or all of the constituent fatty acid.
 7. A composition according toclaim 6, characterized in that the arachidonic acid content of thetriglycerides including triglycerides containing arachidonic acid aspart or all of the constituent fatty acid is at least 10 wt % of thetotal fatty acid in the triglycerides.
 8. A composition according toclaim 6, wherein the triglycerides including triglycerides containingarachidonic acid as part or all of the constituent fatty acid areextracted from microbes belonging to the genus Mortierella.
 9. Acomposition according to claim 6, wherein the triglycerides includingtriglycerides containing arachidonic acid as part or all of theconstituent fatty acid are triglycerides containing no eicosapentaenoicacid or containing no more than 1% eicosapentaenoic acid.
 10. Acomposition with effects of decline prevention, improvement orenhancement of normal responses of cognitive abilities of a healthyperson, which contains triglycerides including at least 5 mole percentof triglycerides with medium-chain fatty acids bound at the1,3-positions and arachidonic acid bound at the 2-position.
 11. Acomposition according to claim 10, wherein the medium-chain fatty acidsare selected from among fatty acids of 6 to 12 carbons.
 12. Acomposition according to claim 11, wherein the medium-chain fatty acidsare selected from among fatty acids of 8 carbons.
 13. A compositionaccording to claim 1 with effects of decline prevention, improvement orenhancement of the normal responses of a healthy person of processingspeed or response speed with respect to events selected from the groupconsisting of auditory stimuli, visual stimuli, olfactory stimuli,gustatory stimuli and somatosensory stimuli, as a cognitive ability. 14.A composition according to claim 1 with effects of decline prevention,improvement or enhancement of the normal response of concentration of ahealthy person with respect to events selected from the group consistingof auditory stimuli, visual stimuli, olfactory stimuli, gustatorystimuli and somatosensory stimuli, as a cognitive ability.
 15. Acomposition according to claim 1 with effects of decline prevention,improvement or enhancement of the normal response of awareness level ofa healthy person, as a cognitive ability.
 16. A composition according toclaim 1 with effects of decline prevention, improvement or enhancementof the normal response of discriminatory ability of a healthy personwith respect to events selected from the group consisting of auditorystimuli, visual stimuli, olfactory stimuli, gustatory stimuli andsomatosensory stimuli, as a cognitive ability.
 17. A compositionaccording to claim 1 with an effect of shortening P300 latency of theevent related potentials of brain (P300), as a response index ofcognitive ability.
 18. A composition according to claim 1 with an effectof augmenting the P300 amplitude of the event related potentials ofbrain (P300), as a response index of cognitive ability.
 19. Acomposition according to claim 1, wherein the composition is a foodcomposition or pharmaceutical composition.
 20. A food compositioncontaining arachidonic acid and/or a compound with arachidonic acid as aconstituent fatty acid, in an amount such that the daily ingestion foran adult is 0.001-20 g in terms of arachidonic acid.
 21. A foodcomposition according to claim 20, wherein the compound with arachidonicacid as a constituent fatty acid is an arachidonic acid alcohol ester ora triglyceride, phospholipid or glycolipid comprising arachidonic acidas part or all of the constituent fatty acid.
 22. A food compositionaccording to claim 21, wherein the triglyceride containing arachidonicacid as part or all of the constituent fatty acid is a triglyceridehaving medium-chain fatty acids bound at the 1,3-positions andarachidonic acid bound at the 2-position.
 23. A food compositionaccording to claim 22, wherein the medium-chain fatty acids are selectedfrom among fatty acids of 6 to 12 carbons.
 24. A food compositionaccording to claim 23, wherein the medium-chain fatty acids are selectedfrom among fatty acids of 8 carbons.
 25. A food compositioncharacterized in that the composition contains at least 0.001 wt % oftriglycerides having medium-chain fatty acids bound at the 1,3-positionsand arachidonic acid bound at the 2-position.
 26. A food compositionaccording to claim 25, wherein the medium-chain fatty acids are selectedfrom among fatty acids of 6 to 12 carbons.
 27. A food compositionaccording to claim 26, wherein the medium-chain fatty acids are selectedfrom among fatty acids of 8 carbons.
 28. A composition according toclaim 20, wherein the food composition is a functional food, nutritionalsupplement food, special health care food or geriatric food.
 29. Acomposition according to claim 1, which further comprisesdocosahexaenoic acid and/or a compound with docosahexaenoic acid as aconstituent fatty acid.
 30. A composition according to claim 29, whereinthe compound with docosahexaenoic acid as a constituent fatty acid is adocosahexaenoic acid alcohol ester or a triglyceride, phospholipid orglycolipid comprising docosahexaenoic acid as part or all of theconstituent fatty acid.
 31. A composition according to claim 29, whereinthe arachidonic acid/docosahexaenoic acid ratio (by weight) in acombination of the arachidonic acid and docosahexaenoic acid is in therange of 0.1-15.
 32. A composition according to claim 1, whereineicosapentaenoic acid is also present in composition in an amount notexceeding ⅕ of the arachidonic acid in the composition.
 33. A processfor production of a food composition with effects of decline prevention,improvement or enhancement of normal responses of cognitive abilities ofa healthy person, comprising combining arachidonic acid and/or acompound with arachidonic acid as a constituent fatty acid, either aloneor with a food material containing either essentially no or only a traceamount of arachidonic acid.
 34. A method for marketing a compositionwith effects of decline prevention, improvement or enhancement of normalresponses of cognitive abilities of a healthy person containingarachidonic acid and/or a compound with arachidonic acid as aconstituent fatty acid, the method for marketing a composition witheffects of decline prevention, improvement or enhancement of normalresponses of cognitive abilities of a healthy person comprising using apackaging container and/or merchandising tool which indicates that thecomposition and/or components in the composition have effects of declineprevention, improvement or enhancement of normal responses of cognitiveabilities of a healthy person.
 35. A composition with effects of declineprevention, improvement or enhancement of normal responses of cognitiveabilities of a healthy person, comprising composition containingarachidonic acid and/or a compound with arachidonic acid as aconstituent fatty acid which is marketed using a packaging containerand/or merchandising tool for the composition indicating that thecomposition and/or components in the composition have effects of declineprevention, improvement or enhancement of normal responses of cognitiveabilities of a healthy person.